Determination of 4 Kinds of β-Agonists Residues in Braised Meat by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry
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摘要: 建立了一种超高效液相色谱串联质谱(ultra-high performance liquid chromatography-tandem mass spectrometry,UPLC-MS)检测卤肉中4种β-受体激动剂(特布他林、克仑特罗、莱克多巴胺、沙丁胺醇)的方法。样品经β-盐酸葡萄糖醛酸苷酶/芳基硫酸酯酶解后,经SLS固相萃取柱净化。以0.1%甲酸水和乙腈为流动相梯度洗脱,用Thermo Hypersil Gold C18色谱柱分离,ESI+进行多反应监测(MRM),内标法定量。4种β-受体激动剂在浓度为0.5~9.5 μg/L范围内线性关系良好,相关系数(r)均大于0.9988,检出限(LOD)为0.1 μg/kg,定量限(LOQ)为0.3 μg/kg。3个加标水平下(1、5、9 μg/kg)的回收率在87.9%~113.7%之间,RSD为1.48%~9.32%。162批次卤肉样品,有一份卤猪蹄样品检出克伦特罗含量为1.51 μg/kg,莱克多巴胺为3.65 μg/kg;另外一份卤羊肉样品检出克伦特罗含量11.5 μg/kg。本方法快速准确,可用于卤肉中4种β-受体激动剂(特布他林、克仑特罗、莱克多巴胺、沙丁胺醇)的定性、定量检测。
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关键词:
- 超高效液相色谱串联质谱(UPLC-MS) /
- 卤肉 /
- β-受体激动剂 /
- SLS固相萃取柱 /
- 同位素内标
Abstract: An ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS) method was developed for the determination of four β-agonists (terbutaline, clenbuterol, ractopamine, salbutamol) in braised meat. Samples were hydrolyzed by β-glucuronidase and cleaned up by an SLS solid phase extraction column. The separation was performed on a Thermo Hypersil Gold C18 column with a gradient elution of 0.1% formic acid water and acetonitrile as mobile phases, ESI+ was used for multiple response monitoring (MRM) and quantitative analysis by internal standard method. The linear relationship of the four β-agonists was good in the concentration range of 0.5 μg/L to 9.5 μg/L, and the correlation coefficient (r) was greater than 0.9988. The limit of detection (LOD) was 0.1 μg/kg, and the limit of quantitation (LOQ) was 0.3 μg/kg. The recoveries were 87.9%~113.7% and RSDs were 1.48%~9.32% at three spiked levels (1, 5 and 9 μg/kg). In a total of 162 batches of braised meat samples, one sample of braised pig’s trotter was found to contain 1.51 μg/kg of clenbuterol and 3.65 μg/kg of ractopamine. Additionally, another sample of braised lamb was found to contain 11.5 μg/kg of clenbuterol. The method is rapid and accurate, and can be used for qualitative and quantitative determination of four β-agonists (terbutaline, clenbuterol, ractopamine, salbutamol) in braised meat. -
β-受体激动剂又称瘦肉精,是苯乙醇胺类化合物,该类药物可明显提高动物的瘦肉率,因此会被非法添加于畜禽类饲料中[1−3]。当人体食入含有β-受体激动剂的动物组织将会出现心悸、 恶心、肌肉震颤和紧张等症状[4−6]。我国已禁止在畜禽类养殖中使用β-受体激动剂[7],但在利益的驱使下,还是存在β-受体激动剂非法添加的情况[8−10]。
目前对于β-受体激动剂的检测方法,常用酶联免疫法[11−12]、气相色谱-串联质谱法[13−15]和液相色谱-串联质谱法[16−18]等。酶联免疫法假阳性率较高,气相色谱-质谱法需要进行衍生后测定,操作过程复杂,且检测灵敏度低,不适用于多种目标物同时分析[19−21]。相较于其他的检测技术,液相色谱-串联质谱法具有灵敏度高、选择性强、分析速度快和样品使用量少等优点,已经广泛应用于食品污染物监测[10,22−23]。市场监管部门对肉制品的监督检测主要是新鲜肉,一些在饲养中有非法添加的肉就存在制成卤肉售卖的风险。卤料成分极其复杂,因此卤肉基质效应大,容易受到干扰出现假阳性。目前动物源性食品中β-受体激动剂的国标检测方法(GB 31658.22-2022)[24]实验前处理操作过程繁琐,提取的样品溶液非缓冲盐体系,需要两次调节pH,两次氮吹复溶,操作繁琐耗时,不适合大批量样品的检测。近几年文献报道的QuEChERS EMR-Lipid净化快速检测肉制品中β-受体激动剂的方法[25−27],操作简单,但主要是除去样品中的脂肪,无法完全去除卤肉中的杂质,检测卤肉易出现假阳性。
为最大限度地消除基质干扰,本实验采用基质标准曲线,同时采用同位素内标矫正。采用基质标准曲线可使样品溶液和标准溶液有相同的离子化环境,从而减小基质效应[25−26,28],采用与待测化合物相对应的同位素内标校正,同位素内标稀释技术提高了β-受体激动剂定量分析的准确性和回收率,消除基质的干扰[27,29−30]。本文在国标方法GB31658.22-2022的基础上,简化了萃取步骤,且仅需调节pH一次,氮吹复溶一次,建立了将卤肉样品用乙酸钠-醋酸缓冲液提取,仍采用经典的SLS固相萃取柱净化,UPLC-MS测定卤肉中4种β-受体激动剂的方法,为卤肉中β-受体激动剂的检测提供了新的思路。
1. 材料与方法
1.1 材料与仪器
甲醇、乙腈 色谱纯,Thermo公司;甲酸 色谱纯,中国阿拉丁公司;浓氨水 优级纯,沪试公司;乙酸乙酯、盐酸、乙酸钠、醋酸 均为分析纯,均为国药集团化学试剂有限公司;β-葡萄糖醛苷酸酶/芳基硫酸酯酶 硫酸酯酶活性<20000 units/mL,美国CNW;特布他林、克仑特罗、莱克多巴胺、沙丁胺醇、D9-特布他林、D9-克仑特罗、D5-莱克多巴胺、D3-沙丁胺醇 均为TRC加拿大公司,纯度均大于95%;卤肉:卤羊肉、卤牛肉、卤猪肉等 来自于贵州省的九个市州。
SLS固相萃取柱 (500 mg/6 mL) 福州勤鹏生物科技有限公司;QuEChERS dSPE EMR-Lipid、QuEChERS Final Polish 美国安捷伦科技有限公司;Oasis MCX固相萃取柱(500 mg/6 mL)、Oasis HLB固相萃取柱(200 mg/6 mL) Waters公司;Millipore A10超纯水机 美国密理博公司;SCIEX 6500三重四极杆串联质谱仪 美国AB SCIEX公司;3-30KS离心机 德国Sigma公司;DMV-16涡旋混匀仪 广东科寅设备有限公司;N-EVAP氮吹仪 上海安普科技有限公司;SHA-C水浴恒温振荡器 金坛市恒丰仪器厂;F2 pH计 梅特勒-托利多仪器上海有限公司。
1.2 实验方法
1.2.1 标准溶液的配制
1.2.1.1 混合标准储备溶液
分别准确称取0.010 g的特布他林、克仑特罗、莱克多巴胺、沙丁胺醇的标准品于4个10 mL容量瓶中,用甲醇溶解并定容至刻度,摇匀,得到1000 mg/L的各单标储备液。取各单标储备液各0.1 mL于10 mL容量瓶中,用甲醇溶解并定容至刻度,摇匀,得到10 mg/L的混合标准储备溶液。临用前逐级稀释成10 μg/L的混合标准使用液。
1.2.1.2 混合同位素内标储备液
分别准确称取0.010 g的D9-特布他林、D9-克仑特罗、D5-莱克多巴胺、D3-沙丁胺醇标准品于4个10 mL容量瓶中,用甲醇溶解并定容至刻度,摇匀,得到1000 mg/L的各同位素内标储备液。取各同位素内标储备液各0.1 mL于10 mL容量瓶中,用甲醇溶解并定容至刻度,摇匀,得到10 mg/L的混合同位素内标储备溶液。临用前逐级稀释成100 μg/L的混合同位素内标使用液。
1.2.2 样品的前处理
称取绞碎卤肉2.00 g样品于50 mL 离心管中,加100 μL 100 μg/L β-受体激动剂混合同位素内标使用液静置10 min,加10 mL 0.2 mol/L pH5.2的乙酸钠-醋酸缓冲液,均质20 s,加入20 μL β-葡萄糖醛苷酶/芳基硫酸酯酶液,37 ℃水浴振荡器水解16 h,取出冷却,10000 r/min 离心10 min,取上清液,用3.0 mol/L盐酸溶液调节pH至2.0±0.1(pH测定仪),10000 r/min 离心10 min,上清液待净化。
SLS固相萃取柱依次用5 mL水、5 mL甲醇、5 mL水、5 mL 40 mmol/L的盐酸溶液活化后,取5 mL上述样品上清液至柱内过柱,然后用5 mL水、5 mL甲醇依次淋洗除杂,抽干,用10 mL 5%氨化乙酸乙酯洗脱,收集洗脱液于10 mL聚丙烯离心管中,50 ℃水浴中氮吹干。初始流动相复溶,涡旋混匀,0 ℃ 7000 r/min离心5 min,上清液进UPLC-MS测定。
1.2.3 仪器条件
1.2.3.1 色谱条件
色谱柱:Thermo Hypersil Gold C18 (100 mm×2.1 mm,1.9 μm);柱温:35 ℃;进样量:5 μL;流速0.3 mL/min;流动相A为0.1%甲酸-水;流动相B为乙腈;梯度洗脱条件:0~0.5 min 95%A,0.5~8.0 min 95%的A线性变化为40%,8.0~9.0 min 40%A,9.1~13.0 min 95%A。
1.2.3.2 质谱条件
离子源:ESI+;离子化电压:4500 V;气帘气压强:35 psi;喷雾气压强:50 psi;辅助加热气压强:50 psi;碰撞气:Medium。多反应监测(MRM)模式进行采集。
1.3 数据处理
UPLC-MS配有SCIEX OS数据处理软件建立标准曲线、计算结果,采用Microsoft Excel处理数据及绘制图表。谱图为Photoshop作图软件绘制。本文净化柱的选择和回收率均采用6个平行样品计算。
2. 结果与分析
2.1 液相条件的确定
在电喷雾离子源正离子模式下,流动相中加入适量甲酸可增强待测组分离子化效率,提高检测响应强度,且乙腈比甲醇有更强的洗脱能力,因此选择乙腈-0.1%甲酸水作为流动相,4种β-受体激动剂的色谱峰形和响应信号的强度也较好。调整洗脱程序,使各组分分离,优化后的色谱条件见1.2.3.1,优化后的谱图如图1所示。
2.2 质谱条件的确定
选择ESI+模式对目标物进行质谱扫描。将质量浓度为50 μg/L的各标准液进入质谱,通过Q1扫描找到各目标物的母离子,再对母离子施加一定的碰撞能量进行Product Ion(MS2)扫描,获得各自的二级碎片离子,选择2个信号较强且干扰小的碎片离子与其母离子组成监测离子对。优化碰撞能量、锥孔电压等各项质谱参数,建立MRM的检测模式。最终确定的离子对信息及质谱参数如表1所示。
表 1 质谱参数Table 1. Mass spectrometric parameters化合物名称 母离子(m/z) 子离子(m/z) 碰撞能量(eV) 去簇电压(V) 扫描时间(ms) 克仑特罗 277.1 203.1* 23 32 100 259.2 16 31 100 D9-克仑特罗 286.2 204.1 23 91 100 特布他林 226.2 152.3* 23 31 100 125.1 34 27 100 D9-特布他林 235.2 153.2 23 85 100 莱克多巴胺 302.2 164.3* 23 37 100 284.4 17 37 100 D5-莱克多巴胺 307.2 165.3 23 52 100 沙丁胺醇 240.1 148.2* 26 35 100 222.1 15 34 100 D3-沙丁胺醇 243.2 151.1 27 53 100 注:*为定量离子。 2.3 净化条件的选择
本课题组李磊等[31]已确证QuEChERS EMR-Lipid净化HPLC-MS测定猪肉猪肝中β-受体激动剂简便可行,在实验中发现此净化方法测定卤肉中β-受体激动剂,高检出率和同一样品同时检出多种β-受体激动剂,数据可疑,可能出现假阳性。如图2所示,同一卤肉样品经SLS柱净化后上机为a图,克伦特罗和莱克多巴胺均未检出。QuEChERS EMR-Lipid净化后上机如b图,在5.28 min处出峰,说明莱克多巴胺有检出;在5.74 min处出峰,说明克伦特罗有检出。采用国标法对这一卤肉样品进行净化复检,克伦特罗和莱克多巴胺均未检出。结果证实QuEChERS EMR-Lipid净化虽然快速简便,但清除不干净卤肉样品中杂质,容易导致假阳性结果,不适合卤肉样品的净化。QuEChERS EMR-Lipid是一种分散固相萃取技术,原理是特殊聚合物基质EMR专门吸附脂质中C5及以上的碳链,对脂质具有非常强的选择吸附性,主要用途是在前处理过程中去除食品基质中的脂质。而经典的SLS柱是利用选择性吸附与选择性洗脱的液相色谱法分离原理,是使液体样品溶液通过吸附剂,保留其中被测物质,再选用适当强度溶剂冲去杂质,然后用少量溶剂迅速洗脱被测物质,从而能更加完全地清除杂质,更适用于复杂基质。
为了使目标化合物的定量分析更加准确,采用的净化柱要能够降低或消除样品基质带来的干扰。肉类中β-受体激动剂的前处理多采用PXC柱、HLB柱、SLS柱、MCX柱、QuEChERS EMR-Lipid等净化,本研究排除了QuEChERS EMR-Lipid,又对比了SLS柱、HLB柱和MCX柱净化的效果。对加标量5.0 μg/kg空白卤牛肉的净化效率,平行测定6次,对比其平均回收率,RSD在1.23%~7.92%之间,结果见图3。
结果显示:HLB柱净化,4种β-受体激动剂的回收率都不高,这是因为HLB柱是共价键结合分配原理,不适用于β-受体激动剂这类中等强极性化合物[32]。SLS柱和MCX柱净化均为强阳离子交换柱,但SLS柱的回收率为89.2%~97.9%优于MCX柱的83.2%~90.4%(详细结果见图3),因此选择SLS柱作为净化柱。
2.4 标准曲线、检出限与定量限
取适量的10 μg/L的混合标准使用液和100 μg/L的混合同位素内标使用液,用空白基质溶液配制成含混合内标5 μg/L的0.5、1.0、2.0、4.0、6.0、8.0、9.5 μg/L基质匹配标准系列溶液。以系列标准溶液中分析物的浓度(μg/L)与对应的同位素内标的峰面积比值绘制校正曲线,按内标法计算试样中β-受体激动剂含量。以3倍信噪比计算方法的检出限(3S/N),以10倍信噪比计算方法的定量限(10S/N),结果见表2。
表 2 β-激动剂线性方程、检出限和定量限Table 2. Linear equation, detection limit and quantitation limit of β-agonists化合物 线性回归方程 相关系数 检出限
(μg/kg)定量限
(μg/kg)克仑特罗 y=0.20865x−0.00440 0.99918 0.1 0.3 特布他林 y=0.29622x+0.02664 0.99953 0.1 0.3 莱克多巴胺 y=0.36347x−0.01383 0.99948 0.1 0.3 沙丁胺醇 y=0.21278x−0.01485 0.99882 0.1 0.3 由表2可知4种β-受体激动剂在0.5~9.5 μg/L浓度范围内r值>0.9988,计算特布他林、克仑特罗、莱克多巴胺、沙丁胺醇的检出限均为0.1 μg/kg,定量限为0.3 μg/kg。与部分已有方法[21,29]相比检出限和定量限更低,定量更为精准可靠。
2.5 回收率与精密度
在2.0 g空白卤肉样品中加入1、5、9 μg/kg的3个不同浓度水平的4种β-受体激动剂标准溶液,同时加入含有和标准曲线相同浓度的内标溶液,每个浓度水平平行测定6次,计算回收率及测定值的相对标准偏差(RSD),结果见表3。
表 3 β-激动剂回收率与精密度结果Table 3. Spiked recoveries and RSDs of β-agonists化合物 加标浓度(μg/g) 测定值(μg/g) 回收率(%) RSD(%) 克仑特罗 1.00 0.90 90.2 8.47 5.00 4.84 96.9 1.48 9.00 8.99 99.9 2.33 特布他林 1.00 0.90 90.3 9.32 5.00 5.69 113.7 8.85 9.00 9.43 104.8 4.00 莱克多巴胺 1.00 0.93 93.3 7.50 5.00 4.39 87.9 6.54 9.00 8.12 90.2 7.89 沙丁胺醇 1.00 0.99 98.7 9.02 5.00 4.69 93.9 5.85 9.00 8.44 93.8 4.85 由表3可知,4种β-受体激动剂的回收率在87.9%~113.7%之间,RSD在1.48%~9.32%之间,表明方法回收率和精密度良好。
2.6 实际样品测定
从贵州省的九个市州每个市(州)采集18份样本,其中18份样本要求区域内的每个县(区)至少采集2份,所有采集的样本均为当地集市随机抽取。用SLS固相萃取柱净化,超高效液相色谱-串联质谱法对采集的162批次卤肉试样(有卤羊肉,卤牛肉,卤猪肉等)中的4种β-受体激动剂进行检测,结果有2份阳性样品。其中采自遵义市农贸市场的一份卤猪蹄样品克伦特罗含量为1.51±0.16 μg/kg,莱克多巴胺为3.65±0.28 μg/kg;另外一份采自遵义市农贸市场的卤羊肉样品克伦特罗含量为11.5±0.29 μg/kg。用国标方法对阳性样本进行复检,卤猪蹄样品克伦特罗含量为1.56±0.07 μg/kg,莱克多巴胺为3.51±0.15 μg/kg;卤羊肉样品克伦特罗含量为11.7±0.20 μg/kg。食品中兽药最大残留限量标准GB 31650-2019和食品中41种兽药最大残留限量GB 31650.1-2022中克伦特罗、特布他林、莱克多巴胺、沙丁胺醇都是禁用药物,而本实验在卤肉中检出,已有食品风险安全隐患,相关部门应引起重视。
3. 结论
本研究建立了SLS固相萃取柱净化,超高效液相色谱-串联质谱法检测卤肉中克伦特罗、特布他林、莱克多巴胺、沙丁胺醇4种β-受体激动剂残留的方法,并对仪器条件以及前处理方法进行了优化。该方法能有效地除去基质干扰,检出限为0.1 μg/kg,定量限为0.3 μg/kg,加标回收率在87.9%~113.7%之间,相对标准偏差1.48%~9.32%。该方法方便快速、准确可靠,满足实际样品检测要求,可为卤肉中β-受体激动剂类化合物兽药残留的监测提供技术支撑,有助于卤肉中β-受体激动剂类化合物兽药残留的市场监管。
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表 1 质谱参数
Table 1 Mass spectrometric parameters
化合物名称 母离子(m/z) 子离子(m/z) 碰撞能量(eV) 去簇电压(V) 扫描时间(ms) 克仑特罗 277.1 203.1* 23 32 100 259.2 16 31 100 D9-克仑特罗 286.2 204.1 23 91 100 特布他林 226.2 152.3* 23 31 100 125.1 34 27 100 D9-特布他林 235.2 153.2 23 85 100 莱克多巴胺 302.2 164.3* 23 37 100 284.4 17 37 100 D5-莱克多巴胺 307.2 165.3 23 52 100 沙丁胺醇 240.1 148.2* 26 35 100 222.1 15 34 100 D3-沙丁胺醇 243.2 151.1 27 53 100 注:*为定量离子。 表 2 β-激动剂线性方程、检出限和定量限
Table 2 Linear equation, detection limit and quantitation limit of β-agonists
化合物 线性回归方程 相关系数 检出限
(μg/kg)定量限
(μg/kg)克仑特罗 y=0.20865x−0.00440 0.99918 0.1 0.3 特布他林 y=0.29622x+0.02664 0.99953 0.1 0.3 莱克多巴胺 y=0.36347x−0.01383 0.99948 0.1 0.3 沙丁胺醇 y=0.21278x−0.01485 0.99882 0.1 0.3 表 3 β-激动剂回收率与精密度结果
Table 3 Spiked recoveries and RSDs of β-agonists
化合物 加标浓度(μg/g) 测定值(μg/g) 回收率(%) RSD(%) 克仑特罗 1.00 0.90 90.2 8.47 5.00 4.84 96.9 1.48 9.00 8.99 99.9 2.33 特布他林 1.00 0.90 90.3 9.32 5.00 5.69 113.7 8.85 9.00 9.43 104.8 4.00 莱克多巴胺 1.00 0.93 93.3 7.50 5.00 4.39 87.9 6.54 9.00 8.12 90.2 7.89 沙丁胺醇 1.00 0.99 98.7 9.02 5.00 4.69 93.9 5.85 9.00 8.44 93.8 4.85 -
[1] XING G, YI C, DOU P Y, et al. Recent progress in the development of β2 adrenergic receptor agonists:A patent review (2015-2020)[J]. Expert Opinion on Therapeutic Patents,2021,31(1/6):1354−3776.
[2] 吴云钊, 张晨, 孙潇, 等. 高效液相色谱-串联质谱/质谱同时测定乳制品中4种β-受体激动剂残留量[J]. 食品科技,2022,47(8):294−301. [WU Y Z, ZHANG C, SUN X, et al. Determination of clenbuterol, salbutamol, ractopamine and terbutaline residues in dairy products by high-performance liquid chromatography-tandem mass spectrometric/mass spectrometry method[J]. Food Science and Technology,2022,47(8):294−301.] doi: 10.3969/j.issn.1005-9989.2022.8.spkj202208045 WU Y Z, ZHANG C, SUN X, et al. Determination of clenbuterol, salbutamol, ractopamine and terbutaline residues in dairy products by high-performance liquid chromatography-tandem mass spectrometric/mass spectrometry method[J]. Food Science and Technology, 2022, 47(8): 294−301. doi: 10.3969/j.issn.1005-9989.2022.8.spkj202208045
[3] 王培龙, 范理, 苏晓鸥, 等. 分子印迹固相萃取-气相色谱-质谱法测定猪尿中4种β-受体激动剂[J]. 分析化学,2012,40(3):470−473. [WANG P L, FAN L, SU X O, et al. Determination of four kinds of β-agonists in swine urine by molecularly imprinted solid phase extraction followed gas chromatography coupled mass spectrometry[J]. Chinese Journal of Analytical Chemistry,2012,40(3):470−473.] WANG P L, FAN L, SU X O, et al. Determination of four kinds of β-agonists in swine urine by molecularly imprinted solid phase extraction followed gas chromatography coupled mass spectrometry[J]. Chinese Journal of Analytical Chemistry, 2012, 40(3): 470−473.
[4] 金玉娥, 郭德华, 郑烨, 等. 液质联用仪测定动物源性食品中11种β2-受体激动剂的研究[J]. 质谱学报,2007(4):193−201. [GIN Y E, GUO D H, ZHEN Y, et al. Determination of 11 β2-agonists in food stuff of animal origin by liquid chromatograph with tandem mass spectrometric detection[J]. Journal of Chinese Mass Spectrometry Society,2007(4):193−201.] doi: 10.3969/j.issn.1004-2997.2007.04.001 GIN Y E, GUO D H, ZHEN Y, et al. Determination of 11 β2-agonists in food stuff of animal origin by liquid chromatograph with tandem mass spectrometric detection[J]. Journal of Chinese Mass Spectrometry Society, 2007(4): 193−201. doi: 10.3969/j.issn.1004-2997.2007.04.001
[5] HOFFMAN R J, HOFFMAN R S, FREYBERG C L, et al. Clenbuterol ingestion causing prolonged tachycardia, hypokalemia, and hypophosphatemia with confirmation by quantitative levels[J]. J Toxicol Clin Toxicol,2001,39(4):339−344.
[6] JUAN C, IGUALADA C, MORAGUES F, et al. Development and validation of a liquid chromatography tandem mass spectrometry method for the analysis of β-agonists in animal feed and drinking water[J]. J Chromatogr A,2010,1217:6061−6068.
[7] 中华人民共和国农业农村部第250号公告. 食品动物中禁止使用的药品及其他化合物清单[Z]. 2020. http://www.moa.gov.cn/nybgb/2020/202002/202004/t20200414_6341556.htm?eqid=e417a3b2000006aa0000000664453f33 [Announcement No. 250th of the Ministry of Agriculture of the People's Republic of China. List of prohibited drugs and other compounds in food animals[Z].2020. http://www.moa.gov.cn/nybgb/2020/202002/202004/t20200414_6341556.htm?eqid=e417a3b2000006aa0000000664453f33] Announcement No. 250th of the Ministry of Agriculture of the People's Republic of China. List of prohibited drugs and other compounds in food animals[Z].2020. http://www.moa.gov.cn/nybgb/2020/202002/202004/t20200414_6341556.htm?eqid=e417a3b2000006aa0000000664453f33
[8] 汪伟, 张坤龙. 20例儿童盐酸克伦特罗食物中毒分析[J]. 安徽预防医学杂志,2011,17(3):219−221. [WANG W, ZHANG K L. Analysis of 20 cases of clenbuterol food poisoning in children[J]. Anhui J Prev Med,2011,17(3):219−221.] WANG W, ZHANG K L. Analysis of 20 cases of clenbuterol food poisoning in children[J]. Anhui J Prev Med, 2011, 17(3): 219−221.
[9] RAHMAN M, HOQUE M, SAZEDUL, et al. Monitoring of pesticide residues from fish feed, fish and vegetables in Bangladesh by GC-MS using the QuEChERS method[J]. Heliyon,2021,7(3):1−8.
[10] FAN S, MIAO H, ZHAO Y, et al. Simultaneous detection of residues of 25 β2-agonists and 23 β-blockers inanimal foods by high-performance liquid chromatography coupled with linear iontrap mass spectrometry[J]. J Agric Food Chem,2013,60(8):1898−1905.
[11] 刘淼苗, 刘啊敏, 陈尔净, 等. 基于二聚人工抗原ELISA检测多残留β2-受体激动剂[J]. 中国计量大学学报,2022,33(1):27−35. [LIU M M, LIU A M, CHEN E J, et al. Detection of β2-agonists multi-residual by ELISA based on dimeric artificial antigen[J]. Journal of China University of Metrology,2022,33(1):27−35.] doi: 10.3969/j.issn.2096-2835.2022.01.005 LIU M M, LIU A M, CHEN E J, et al. Detection of β2-agonists multi-residual by ELISA based on dimeric artificial antigen[J]. Journal of China University of Metrology, 2022, 33(1): 27−35. doi: 10.3969/j.issn.2096-2835.2022.01.005
[12] 李延山, 刘庆伟, 卞大伟, 等. ELISA检测猪尿中β-受体激动剂类药物影响因素研究[J]. 动物医学进展,2018,39(8):78−82. [LI Y S, LIU Q W, BIAN D W, et al. Study on influencing factors for detecting β-receptor agonists in porcine urine by ELISA[J]. Progress in Veterinary Medicine,2018,39(8):78−82.] doi: 10.3969/j.issn.1007-5038.2018.08.017 LI Y S, LIU Q W, BIAN D W, et al. Study on influencing factors for detecting β-receptor agonists in porcine urine by ELISA[J]. Progress in Veterinary Medicine, 2018, 39(8): 78−82. doi: 10.3969/j.issn.1007-5038.2018.08.017
[13] 朱坚, 李波, 方晓明, 等. 气相色谱质谱法测定肝、肾和肉中11种β-受体激动剂残留量[J]. 质谱学报,2005(3):129−137. [ZHU J, LI B, FANG X M, et al. Analysis of 11 β-agonist residues in liver, kidney and meat by gas chromatography-mass spectrometry[J]. Journal of Chinese Mass Spectrometry Society,2005(3):129−137.] doi: 10.3969/j.issn.1004-2997.2005.03.001 ZHU J, LI B, FANG X M, et al. Analysis of 11 β-agonist residues in liver, kidney and meat by gas chromatography-mass spectrometry[J]. Journal of Chinese Mass Spectrometry Society, 2005(3): 129−137. doi: 10.3969/j.issn.1004-2997.2005.03.001
[14] LIU W, ZHANG L, WEI Z, et al. Analysis of β-agonists and β-blockers in urine using hollow fibre-protected liquid-phase microextraction with in situ derivatization followed by gas chromatography/mass spectrometry[J]. Journal of Chromatography A,2009,1216(28):5340−5346.
[15] 岳韩笑, 雷雯, 杜晓宁, 等. 同位素稀释-气相色谱-串联质谱法测定猪肉中残留的4种β-受体激动剂[J]. 质谱学报,2018,39(1):61−68. [YUE H X, LEI W, DU X N, et al. Multi-residue analysis of 4 β-agonists in pork using isotope dilution gas chromatography-tandem mass spectrometry[J]. Journal of Chinese Mass Spectrometry Society,2018,39(1):61−68.] doi: 10.7538/zpxb.2016.0123 YUE H X, LEI W, DU X N, et al. Multi-residue analysis of 4 β-agonists in pork using isotope dilution gas chromatography-tandem mass spectrometry[J]. Journal of Chinese Mass Spectrometry Society, 2018, 39(1): 61−68. doi: 10.7538/zpxb.2016.0123
[16] MEHRABIAN M, NOROOZIAN E, MAGHSOUDI S H. Preparation and application of Fe3O4@SiO2@poly(o-phenylenediamine)nanoparticles as a novel magnetic sorbent for the solid-phase extraction of tellurium in water samples and its determination by ET-AAS[J]. Microchem J,2021,165(6):104−106.
[17] MARIA A H, APPU R. Basic pharmacology:Understanding drug actions and reactions, second edition[M]. CRC Press:2020.
[18] 文娜, 毛琼玲, 左书瑞, 等. UPLC-MS/MS测定畜禽肉中利巴韦林药物与β-受体激动剂残留量[J]. 安徽农业科学,2022,50(24):199−203. [WEN N, MAO Q L, ZUO S D, et al. Determination of ribavirin and β-receptor agonist residues in livestock and poultry meat by UPLC-MS/MS[J]. Journal of Anhui Agricultural Sciences,2022,50(24):199−203.] doi: 10.3969/j.issn.0517-6611.2022.24.048 WEN N, MAO Q L, ZUO S D, et al. Determination of ribavirin and β-receptor agonist residues in livestock and poultry meat by UPLC-MS/MS[J]. Journal of Anhui Agricultural Sciences, 2022, 50(24): 199−203. doi: 10.3969/j.issn.0517-6611.2022.24.048
[19] 王远, 邢丽杰, 郝家勇, 等. 高效液相色谱-串联质谱法测定卤肉中3种β-受体激动剂残留[J]. 食品科学,2013,34(8):216−219. [WANG Y, XING L J, HAO J Y, et al. Determination of β-agonists in cooked meat by HPLC-MS/MS[J]. Food Science,2013,34(8):216−219.] doi: 10.7506/spkx1002-6630-201308046 WANG Y, XING L J, HAO J Y, et al. Determination of β-agonists in cooked meat by HPLC-MS/MS[J]. Food Science, 2013, 34(8): 216−219. doi: 10.7506/spkx1002-6630-201308046
[20] ZHANG J Y, WEI Y H, QIU S Y, et al. A highly selective and simple fluorescent probe for salbutamol detection based on thioglycolic acid-capped CdTe quantum dots[J]. Spectrochim Acta Part A:Mol Biomol Spectrosc,2021,247(2):107−119.
[21] 李丽珍, 吴学贵, 李小梅, 等. 高效液相色谱-串联质谱法测定猪肉中8种β-受体激动剂残留[J]. 食品安全质量检测学报,2020,11(5):1642−1650. [LI L Z, WU X G, LI X M, et al. Determination of eight kinds of β-agonist residues in pork by high performance liquid chromatography-tandem mass spectrometry[J]. Journal of Food Safety & Quality,2020,11(5):1642−1650.] doi: 10.19812/j.cnki.jfsq11-5956/ts.2020.05.053 LI L Z, WU X G, LI X M, et al. Determination of eight kinds of β-agonist residues in pork by high performance liquid chromatography-tandem mass spectrometry[J]. Journal of Food Safety & Quality, 2020, 11(5): 1642−1650. doi: 10.19812/j.cnki.jfsq11-5956/ts.2020.05.053
[22] 王莉莉, 张楠, 刘平, 等. 通过式固相萃取柱结合QuEChERS前处理技术-液相色谱串联质谱法快速测定熟肉食品中4种β2-受体激动剂残留[J]. 食品安全质量检测学报,2021,12(9):3771−3776. [WANG L L, ZHANG N, LIU P, et al. Rapid determination of 4 kinds of β2-agonist residues in cooked meat by pass-through solid phase extraction column combined with QuEChERS pretreatment technique-liquid chromatography tandem mass spectrometry[J]. Journal of Food Safety & Quality,2021,12(9):3771−3776.] WANG L L, ZHANG N, LIU P, et al. Rapid determination of 4 kinds of β2-agonist residues in cooked meat by pass-through solid phase extraction column combined with QuEChERS pretreatment technique-liquid chromatography tandem mass spectrometry[J]. Journal of Food Safety & Quality, 2021, 12(9): 3771−3776.
[23] BING S, JIA X, JING Z, et al. Multi-residual analysis of 16 β-agonists in pig liver, kidney and muscle by ultra performance liquid chromatography tandem mass spectrometry[J]. Food Chemistry,2009,114(3):1115−1121.
[24] 中华人民共和国农村农业部, 中华人民共和国国家卫生健康委员会, 国家市场监督管理总局. 食品安全国家标准 动物性食品中β-受体激动剂残留量的测定 液相色谱-串联质谱法:GB31658.22-2022[S]. 北京:中国农业出版社, 2022. [Ministry of Rural Agriculture of the People's Republic of China, National Health Commission of the People's Republic of China, State Administration of Market Supervision and Administration. National food safety standard-Determination of B-agonists residues in animal derived food by liquid chromatography-tandem mass spectrometric method:GB31658.22-2022[S]. Beijing:China Agricultural Press, 2022.] Ministry of Rural Agriculture of the People's Republic of China, National Health Commission of the People's Republic of China, State Administration of Market Supervision and Administration. National food safety standard-Determination of B-agonists residues in animal derived food by liquid chromatography-tandem mass spectrometric method: GB31658.22-2022[S]. Beijing: China Agricultural Press, 2022.
[25] 李婧妍, 韩波, 安乐, 等. QuEChERS前处理结合HPLC-MS/MS同时测定原料乳中30种兽药残留[J]. 食品研究与开发,2022,43(9):176−185. [LI J Y, HAN B, AN L, et al. Simultaneous determination of residues of 30 veterinary drugs in raw milk by QuEChERS and HPLC-MS/MS[J]. Food Research and Development,2022,43(9):176−185.] doi: 10.12161/j.issn.1005-6521.2022.09.024 LI J Y, HAN B, AN L, et al. Simultaneous determination of residues of 30 veterinary drugs in raw milk by QuEChERS and HPLC-MS/MS[J]. Food Research and Development, 2022, 43(9): 176−185. doi: 10.12161/j.issn.1005-6521.2022.09.024
[26] 张科明, 梁飞燕, 邓鸣, 等. QuEChERS结合液相色谱-串联质谱法快速测定猪肉中多类兽药残留[J]. 色谱,2016,34(9):860−867. [ZHANG K M, LIANG F Y, DENG M, et al. Multiresidue determination of veterinary drugs in porcine muscle by QuEChERS and liquid chromatography tandem mass spectrometry[J]. Chinese Journal of Chromatography,2016,34(9):860−867.] doi: 10.3724/SP.J.1123.2016.06010 ZHANG K M, LIANG F Y, DENG M, et al. Multiresidue determination of veterinary drugs in porcine muscle by QuEChERS and liquid chromatography tandem mass spectrometry[J]. Chinese Journal of Chromatography, 2016, 34(9): 860−867. doi: 10.3724/SP.J.1123.2016.06010
[27] 毛锐, 林浩, 刘川, 等. QuEChERS EMR-Lipid净化结合同位素稀释-超高效液相色谱-串联质谱法同时测定畜肉中17种β-受体激动剂[J]. 食品工业科技,2023,44(15):320−328. [MAO R, LIN H, LIU C, et al. Simultaneous determination of 17 β-receptor agonists in meat by QuEChERS EMR-Lipid with isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry[J]. Science and Technology of Food Industry,2023,44(15):320−328.] MAO R, LIN H, LIU C, et al. Simultaneous determination of 17 β-receptor agonists in meat by QuEChERS EMR-Lipid with isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry[J]. Science and Technology of Food Industry, 2023, 44(15): 320−328.
[28] 赵鹏, 常靖, 吴小军, 等. 超高效液相色谱-串联质谱检测干血斑中29种β2-受体激动剂[J]. 分析测试学报,2022,41(2):227−233. [ZHAO P, CHANG J, WU X J, et al. Determination of 29 β2-agonists in dried blood spots by ultra-high performance liquid chromatography-tandem mass spectrometry[J]. Journal of Instrumental Analysis,2022,41(2):227−233.] ZHAO P, CHANG J, WU X J, et al. Determination of 29 β2-agonists in dried blood spots by ultra-high performance liquid chromatography-tandem mass spectrometry[J]. Journal of Instrumental Analysis, 2022, 41(2): 227−233.
[29] 李艳明, 王坤, 朱富强, 等. 通过式固相萃取-超高效液相色谱-串联质谱法测定猪肉中多种受体激动剂药物残留[J]. 食品工业科技,2021,42(14):264−271. [LI Y M, WANG K, ZHU F Q, et al. Determination of receptor agonist residues in pork by solid-phase extraction coupled with ultra-performance liquid chromatography-tandem mass spectrometry[J]. Science and Technology of Food Industry,2021,42(14):264−271.] doi: 10.13386/j.issn1002-0306.2020090182 LI Y M, WANG K, ZHU F Q, et al. Determination of receptor agonist residues in pork by solid-phase extraction coupled with ultra-performance liquid chromatography-tandem mass spectrometry[J]. Science and Technology of Food Industry, 2021, 42(14): 264−271. doi: 10.13386/j.issn1002-0306.2020090182
[30] 吴永宁, 苗虹, 范赛, 等. 高效液相色谱-线性离子阱质谱法测定畜禽肌肉中β2-受体激动剂及β-阻断剂类药物残留[J]. 中国科学(B辑:化学),2009,39(8):774−784. [WU YN, MIAO H, FAN S, et al. Determination of β2-agonist and-blocker residues in livestock and poultry muscle by HIGH performance liquid chromatography-linear ion trap mass spectrometry[J]. Sci China(Ser B:Chem),2009,39(8):774−784.] WU YN, MIAO H, FAN S, et al. Determination of β2-agonist and-blocker residues in livestock and poultry muscle by HIGH performance liquid chromatography-linear ion trap mass spectrometry[J]. Sci China(Ser B:Chem), 2009, 39(8): 774−784.
[31] 李磊, 李海畅, 高婧, 等. QuEChERS EMR-Lipid-LC/MS/MS测定8种β-受体激动剂[J]. 食品研究与开发,2016,37(9):178−182. [LI L, LI H C, GAO J, et al. Determination of 8 β-agonists by QuEChERS EMR-Lipid-LC/MS/MS[J]. Food Research and Development,2016,37(9):178−182.] doi: 10.3969/j.issn.1005-6521.2016.09.042 LI L, LI H C, GAO J, et al. Determination of 8 β-agonists by QuEChERS EMR-Lipid-LC/MS/MS[J]. Food Research and Development, 2016, 37(9): 178−182. doi: 10.3969/j.issn.1005-6521.2016.09.042
[32] 史娜, 郝杰, 姜洁, 等. 超高效液相色谱-串联质谱法同时检测动物源性食品中残留的35种β-受体激动剂和11种β-受体阻断剂[J]. 分析仪器,2018(1):20−31. [SHI N, HAO J, JIANG J, et al. Simultaneous determination of 35 β-agonist and 11 β-blocker residues in animal-origin foods by ultra high performance liquid chromatography-tandem mass spectrometry[J]. Analytical Instrumentation,2018(1):20−31.] doi: 10.3969/j.issn.1001-232x.2018.01.004 SHI N, HAO J, JIANG J, et al. Simultaneous determination of 35 β-agonist and 11 β-blocker residues in animal-origin foods by ultra high performance liquid chromatography-tandem mass spectrometry[J]. Analytical Instrumentation, 2018(1): 20−31. doi: 10.3969/j.issn.1001-232x.2018.01.004